Deoxynivalenol and nivalenol in commercial wheat grain related to Fusarium head blight epidemics in southern Brazil

A three-year (2006-2008) survey on commercial wheat grain was conducted aimed at quantifying the intensity of Fusarium head blight epidemics related to kernel quality and levels of deoxynivalenol (DON) and nivalenol (NIV). Grain samples, obtained from 38 municipalities throughout the state of Rio Grande do Sul, Brazil, were assessed visually for Fusarium-damaged kernels (FDK) and chemically using liquid chromatography-mass spectrometry (LC-MS/MS). Overall FDK mean levels were 15.5%, not differing among the years. Co-contamination was predominant (59/66) across samples and overall mean levels of DON and NIV were 540 and 337 μg/kg, respectively. When the levels of both mycotoxins were added together (DON + NIV), a higher correlation with FDK was found (R = 0.36, P < 0.01), compared to single toxin data. For the first time, the presence of NIV in levels comparable to DON is reported from a multi-year regional epidemiological survey in the country which should be of concern to the small grains industry.     

Effects of Fusarium infection on the amino acid composition of winter wheat grain

Winter wheat (susceptible cultivar Ritmo) was grown in 2006 near Kiel and in 2007 near Heide in northern Germany. Plants were inoculated at anthesis using a Fusarium graminearum macroconidial suspension. The percentage of Fusarium-damaged kernels (FDK) ranged from 0 ± 2% to 28 ± 2%. The contents of the Fusarium mycotoxin deoxynivalenol (DON) and wheat amino acids were determined in the grain. Levels of the amino acids alanine, lysine, and tyrosine increased with the percentage of FDK or DON contents whereas glutamic acid contents decreased. Aspartic acid and threonine were not related to the percentage of FDK or DON contents. Effects of Fusarium infection on other amino acids were significant only at the sampling site with the higher degree of Fusarium-damage. Interestingly, those amino acids that increased consistently and significantly with the degree of Fusarium-damage are derived from phosphoenolpyruvate or pyruvate, suggesting that pathogen-induced changes in the glycolytic input for amino acid biosynthesis play a significant role for the amino acid composition of Fusarium-damaged winter wheat grain. On average, amino acid contents decreased by 0.13% compared to the amino acid content of sound kernels upon an increase of 1% of FDK.     

Determination of total trans fat content in Pakistani cereal-based foods by SB-HATR FT-IR spectroscopy coupled with partial least square regression

The purpose of this study was to determine the total trans fat content from the Fourier transform infrared (FT-IR) spectra of neat fats extracted from cereal-based foods using a single-bounce horizontal attenuated total reflectance (SB-HATR) sampling accessory. Partial least squares (PLS) regression was employed for the development of calibration models. A trans fat calibration curve was prepared by adding trielaidine gravimetrically to trans free canola oil. Regression of the FT-IR/PLS-predicted trans contents was 0.999 with a root-mean-square error of calibration (RMSEC) and root-mean-square error of prediction (RMSEP) 0.07% and 0.07%, respectively. Amongst the samples examined, the fat content was in the range of 8.2–27.2%, whilst most samples contained more than 10% trans fat; the highest trans fat content was 16.3%. The FT-IR results were comparable with gas chromatography (GC).     

禾谷镰刀菌脱氧雪腐镰刀菌烯醇生物合成及其分子调控研究进展

禾谷镰刀菌是感染小麦、玉米等主要粮食作物的一种重要的病原真菌,在引发严重小麦赤霉病害的同时,可分泌产生脱氧雪腐镰刀菌烯醇（DON）,污染食品后不易被清除,严重威胁人和动物的健康。禾谷镰刀菌产DON的生物合成路径及分子调控机制一直是国内外的研究热点、焦点和难点,也是防控DON的基础。本文综述了禾谷镰刀菌DON生物合成过程中涉及产毒基因簇TRI的各个基因、编码蛋白及其生物学功能,以及目前对禾谷镰刀菌中对呕吐毒素生物合成调控机制的研究进展。这些基础信息将直接为安全控制主要粮食及其制品中真菌毒素污染提供有价值的参考资料。     

Functional properties of two low-molecular-weight glutenin subunits carrying additional cysteine residues from hybrid introgression line II-12 derived from Triticum aestivum and Agropyron elongatum

The functional properties of the various alleles of the low-molecular-weight glutenin subunits (LMW-GS) have not been widely addressed, even though they are important in the context of wheat end-use quality improvement. Two LMW-GS alleles introgressed into bread wheat from tall wheatgrass via asymmetric somatic hybridization differ from the norm by encoding either two or three additional cysteine residues. These genes were isolated and expressed heterologously in Escherichia coli to obtain sufficient product amounts for incorporation into the flour of a poor quality cultivar for the purpose of a micro-mixing test. Their effect was to decrease both the time to peak dough resistance and the peak width, while having no effect on either the 8 min curve width or the right of peak slope. Several effects on wheat dough properties were discussed in this paper.     

PorA specific primers for the identification of Campylobacter species in food and clinical samples

Campylobacteriosis is a public health problem with considerable socio-economic impact. As the European Food Safety Authority has emphasized the importance of a surveillance programme for campylobacteriosis, the aim of the present study was the optimization of a specific and sensitive PCR protocol able to detect Campylobacter species responsible for gastrointestinal infections. Raw poultry meat samples were analysed for the presence of Campylobacter sp., by plating onto mCCD (Modified Charcoal-Cefoperazone-Deoxycholate) Agar and Campylobacter Selective Preston Agar and using four sets of species-specific primers for Campylobacter jejuni, Campylobacter coli, Campylobacter upsaliensis and Campylobacter lari designed to bridge the porA gene. The resulting primers demonstrated a sensitivity of 0.01 ng/μl for the C. coli-specific, C. lari-specific, and C. upsaliensis-specific primer sets and 0.5 ng/μl for the C. jejuni-specific primer sets using DNA from pure cultures. Non-specific amplification of non-target DNA was not observed indicating excellent specificity. The primers were useful for the analyses of poultry meat samples both for direct plating onto mCCDA, and for DNA extracted directly from the cells grown for 48 h in Preston enrichment broth. The sets of primers were also useful when used for species identification of human isolates.     

Antioxidant and free radical scavenging activities of whole wheat and milling fractions

The effects of milling on the phenolic content and antioxidant capacity of two wheat cultivars, namely CWAD (Canadian Western Amber Durum; Triticum turgidum L. var. durum) and CWRS (Canadian Western hard red spring; Triticum aestivum L.) were studied. The milling of wheat afforded several fractions, namely bran, flour, shorts and feed flour. In addition, semolina was the end-product of durum wheat milling. Among different milling fractions the bran had the highest phenolic content while the endosperm possessed the lowest amount and this was also reflected in free radical and reactive oxygen species (ROS) scavenging capacity, reducing power and iron (II) chelation capacity of different milling fractions in the two cultivars. This study demonstrated the importance of bran in the antioxidant activity of wheat, hence consumption of whole wheat grain may render beneficial health effects.     

An investigation of the bacteriocinogenic potential of lactic acid bacteria associated with wheat (Triticum durum) kernels and non-conventional flours

One hundred and thirty-seven lactic acid bacteria (LAB), previously isolated from wheat (Triticum durum) grains and non-conventional flour samples, were tested for the production of antibacterial substances. A total of 16 strains (5 Enterococcus faecium, 5 Enterococcus mundtii, 4 Pediococcus pentosaceus, 1 Lactobacillus coryniformis and 1 Lactococcus garvieae) were found to inhibit the growth of Listeria innocua. The antibacterial activities were preliminarily investigated for their general behaviour with proteolytic (proteinase K, protease B and trypsin), amylolytic (α-amylase) and lipolytic (lipase) enzymes, after heat treatment, and exposure to different pHs and ethanol concentrations. Bacteriocin-like inhibitory substances (BLIS) were also characterized for their inhibition spectra against non-pathogenic and pathogenic food-associated and human pathogenic bacteria. LAB showing the best characteristics in terms of inhibition spectrum, inhibition activity and mode of action (bactericidal) belonged to the species Ent. mundtii. The high percentage (11.68%) of BLIS-producing strains detected confirmed previous observations that raw materials may harbour higher numbers of bacteriocinogenic LAB than fermented foods.     

Fast,clean, low-cost screening of cadmium and lead in the mussel Mytilus galloprovincialis Lmk. by visible spectroscopy and partial least squares regression

The potential of visible (VIS) spectroscopy for screening cadmium and lead contents in the mussel Mytilus galloprovincialis Lmk. was assessed. The spectra of the samples were recorded (400–700 nm). The Cd and Pb contents were determined by graphite furnace atomic absorption spectroscopy, and were regressed against different spectral transformations by modified partial least square (PLSm) regression. Coefficients of determination in the cross-validation (Cd = 0.95; Pb = 0.77) were indicative of equations with excellent and good quantitative information, respectively. The standard deviation to standard error of cross-validation ratio (Cd = 4.43; Pb = 2.10) showed sufficient predictive accuracy for the equations to be used for screening purposes. PLSm loading plots corresponding to the first terms of the equations showed that chromophores influenced them significantly. This pioneering use of the VIS spectrum to predict Cd and Pb in mussels represents an important saving in time and cost of analysis in comparison to other methods.     

A novel approach to authenticity control of whole grain durum wheat (Triticum durum Desf.) flour and pasta,based on analysis of alkylresorcinol composition

Since durum wheat is ∼20% more expensive than common wheat and considered of superior quality for the manufacture of pasta products, efficient methods for the detection of accidental or intentional admixtures of common wheat to durum wheat products are required. This paper describes a novel approach for the detection and quantification of whole grain common wheat adulteration in whole grain durum flour and dried pasta. We found that differences in the C17:0 to C21:0 alkylresorcinol homologue ratios between the two cereal species may serve as a suitable tool for whole grain durum product authentification. To detect and estimate adulteration, the C17:0/C21:0 ratios of flour and pasta admixtures with added whole grain flour of common wheat were analysed. A linear relationship between C17:0/C21:0 ratios and level of admixture in pasta samples showed that adulteration can be estimated within the range of 5–100% of admixture. Furthermore, di- and triunsaturated as well as oxygenated alk(en)ylresorcinols are reported to occur in Triticum durum Desf. for the first time.     

Inactivation of Escherichia coli by ozone treatment of apple juice at different pH levels

This research investigated the efficacy of gaseous ozone on the inactivation of Escherichia coli ATCC 25922 and NCTC 12900 strains in apple juice of a range of pH levels, using an ozone bubble column. The pH levels investigated were 3.0, 3.5, 4.0, 4.5 and 5.0. Apple juice inoculated with E. coli strains (10⁶ CFU/mL) was treated with ozone gas at a flow rate of 0.12 L/min and ozone concentration of 0.048 mg/min/mL for up to 18 min. Results show that inactivation kinetics of E. coli by ozone were affected by pH of the juice. The ozone treatment duration required for achieving a 5-log reduction was faster (4 min) at the lowest pH than at the highest pH (18 min) studied. The relationship between time required to achieve 5 log reduction (t_5d) and pH for both strains was described mathematically by two exponential equations. Ozone treatment appears to be an effective process for reducing bacteria in apple juice and the required applied treatment for producing a safe apple juice is dependant on its acidity level.     

Rapid method for controlling the correct labeling of products containing common octopus (Octopus vulgaris) and main substitute species (Eledone cirrhosa and Dosidicus gigas) by fast real-time PCR

The TaqMan real-time PCR has the highest potential for automation, therefore representing the currently most suitable method for screening, allowing the detection of fraudulent or unintentional mislabeling of species. This work describes the development of a real-time polymerase chain reaction (RT-PCR) system for the detection and identification of common octopus (Octopus vulgaris) and main substitute species (Eledone cirrhosa and Dosidicus gigas). This technique is notable for the combination of simplicity, speed, sensitivity and specificity in an homogeneous assay.     

Degradation of ascorbic acid and potassium sorbate by different Lactobacillus species isolated from packed green olives

The aim of this research was to ascertain the lactic acid bacteria responsible for the degradation of ascorbic acid and/or potassium sorbate, isolated from packed green olives where these additives had diminished. A total of 14 isolates were recovered from samples of different green olive containers. According to partial sequencing of the 16S rRNA coding gene, Lactobacillus parafarraginis, Lactobacillus rapi, Lactobacillus pentosus, Lactobacillus paracollinoides, and Pediococcus ethanolidurans were identified. With the exception of L. pentosus and L. paracollinoides, the other species had not been mentioned in table olives before this study. Only three of the 14 isolates metabolized ascorbic acid in MRS broth, and the products from ascorbic acid in modified MRS broth without carbon sources were acetic and lactic acids. Except for the two L. rapi and the two P. ethanolidurans strains, the remaining 10 isolates depleted potassium sorbate added into MRS broth to some extent. The product generated by three of these strains was confirmed to be trans-4-hexenoic acid. The degradation of ascorbate or sorbate by lactic acid bacteria should be taken into account when these additives are used in food products where this group of bacteria may be present.     

Molecular characterization and fumonisin production by Fusarium verticillioides isolated from corn grains of different geographic origins in Brazil

Gibberella moniliformis is most commonly associated with maize worldwide and produces high levels of fumonisins, some of the most agriculturally important mycotoxins. Studies demonstrate that molecular methods can be helpful for a rapid identification of Fusarium species and their levels of toxin production. The purpose of this research was to apply molecular methods (AFLP, TEF-1α partial gene sequencing and PCR based on MAT alleles) for the identification of Fusarium species isolated from Brazilian corn and to verify if real time RT-PCR technique based on FUM1 and FUM19 genes is appropriated to estimate fumonisins B₁ and B₂ production levels. Among the isolated strains, 96 were identified as Fusarium verticillioides, and four as other Fusarium species. Concordant phylogenies were obtained by AFLP and TEF-1α sequencing, permitting the classification of the different species into distinct clades. Concerning MAT alleles, 70% of the F. verticillioides isolates carried the MAT-1 and 30% MAT-2. A significant correlation was observed between the expression of the genes and toxin production r = 0.95 and r = 0.79 (correlation of FUM1 with FB₁ and FB₂, respectively, P < 0.0001); r = 0.93 and r = 0.78 (correlation of FUM19 with FB₁ and FB₂, respectively, P < 0.0001). Molecular methods used in this study were found to be useful for the rapid identification of Fusarium species. The high and significant correlation between FUM1 and FUM19 expression and fumonisins production suggests that real time RT-PCR is suitable for studies considering the influence of abiotic and biotic factors on expression of these genes. This is the first report concerning the expression of fumonisin biosynthetic genes in Fusarium strains isolated from Brazilian agricultural commodity.     

Antioxidant activity and phenolic contents of Olea europaea L. leaves sprayed with different copper formulations

Olive trees (Olea europaea L. Cv. Cobrançosa) from the northeast of Portugal were sprayed with three different copper formulations [bordeaux mixture (copper sulphate + calcium hydroxide − 20% Cu), copper hydroxide (40% Cu) and copper oxychloride (50% Cu)] to control olive fungal diseases. The residues of copper in olive leaves, harvested at different times, were evaluated by atomic absorption spectrometry. At all the collection times, treated olive leaves had significantly higher copper contents, compared to the control. The different copper amounts in pesticide formulations lowered the leaves contents in total phenols and hence their antioxidant properties. Olive leaves sprayed with copper oxychloride possessed the highest copper levels and the lowest content in phenols, which influenced its antioxidant activity (higher EC₅₀ values for reducing power, scavenging effect on DPPH radicals and inhibition of erythrocyte hemolysis). Leaves without copper residues proved to be a good natural source of antioxidants, giving values comparable to the reference compounds.     

Concordance between antioxidant activities and flavonol contents in different extracts and fractions of Cuscuta chinensis

Chinese herbs employed in traditional Chinese medicine (TCM) have been used for centuries in the practice of medicated diet and dietetic therapy. The seed of Cuscuta chinensis Lam. (Convolvulaceae), a commonly used traditional Chinese herb, is frequently added in Chinese cooking and preparation of refreshments, including porridge and alcoholic beverages, to nourish the human body. In the present study, we compared the antioxidant activities of water and ethanol extracts from the seeds C. chinensis and also of its different organic fractions, including n-hexane, ethyl acetate, n-butanol and organic water, by assessing their DPPH (1,1-diphenyl-2-picryl hydrazine) free radical-scavenging, superoxide anion scavenging, anti-superoxide anion formation and anti-lipid peroxidation abilities. The flavonol contents of all test samples were analyzed by high-performance liquid chromatography with an ultraviolet (HPLC–UV) detector. The results showed that there is a direct correlation of the flavonol content with the antioxidant activities from the extracts and fractions of C. chinensis. Moreover, the ethyl acetate fraction demonstrated significantly better and higher antioxidant effects, and also had a higher flavonol content than had the remaining samples (P < 0.05). The water fractions, however, exhibited the weakest antioxidant activity, and had low concentrations of flavonols. Thus, we suggest that the ethanol extract of C. chinensis, but not its water extract, could be used as a dietary nutritional supplement to promote human health and prevent oxidation-related diseases, due to its antioxidant properties.     

Further data on the levels of emerging Fusarium mycotoxins enniatins (A,A1, B,B1), beauvericin and fusaproliferin in breakfast and infant cereals from Morocco

Sixty-eight samples of cereals products, including breakfast cereals (n = 48) and infant cereals (n = 20), purchased from supermarkets and pharmacies in Rabat-Salé area from Morocco were analysed for the determination of six emerging mycotoxins: four enniatins ENs (ENA, ENA1, ENB and ENB1), beauvericin (BEA) and fusaproliferin (FUS). Samples were extracted with a mixture of acetonitrile:water (85:15, v/v), using an Ultra-Turrax® homogeniser. Mycotoxins were then identified and quantified by liquid chromatography (LC) with diode array detection (DAD). Positive samples were confirmed by LC–MS/MS.     

Formation of vinylphenolic pyranoanthocyanins by Saccharomyces cerevisiae and Pichia guillermondii in red wines produced following different fermentation strategies

The strains of two species of yeast, Saccharomyces cerevisiae and Pichia guillermondii, both with high hydroxycinnamate decarboxylase (HCDC) activity (56% and 90% respectively), were used in the fermentation of musts enriched with grape anthocyanins, to favour the formation of highly stable vinylphenolic pyranoanthocyanin pigments. The different strains were used to ferment the must separately, simultaneously, or sequentially, the latter involving an initial period using the yeast with the greatest HCDC activity (P. guillermondii). The must was made from concentrated grape juice diluted to 220 g/l of sugar, and enriched with grape anthocyanins to 100 mg/l and with p-coumaric acid to 120 mg/l. The pH was fixed to 3.5. All 50 ml micro-fermentations were done in triplicate. The development of anthocyanin-3-O-glucoside precursors, the decarboxylation of p-coumaric acid, and the formation of 4-vinylphenol and vinylphenolic pyranoanthocyanin derivatives were studied during the fermentation. The fermentation strategy used and the yeast HCDC activity significantly influenced the formation of vinylphenolic pyranoanthocyanins. The latter molecules were separated, identified, and quantified using high performance liquid chromatograph with diode array detection and electrospray-mass spectrometry (HPLC-DAD-ESI/MS). The volatile compounds profile was screened during fermentation using gas cromatogrphy-flame ionisation detection (GC-FID), in order to detect and quantify the main molecules. The best results were reached with the sequential fermentation (3.74 mg/l of malvidin-3-O-glucoside-4-vinylphenol). This work shows that during mixed or sequential fermentations carried out with non-Saccharomyces or highly fermentative Saccharomyces strains, with high HCDC activity, the content of stable pigments can be increased without sensorial modifications.